scholarly journals Sperm DNA fragmentation in cryopreserved samples from subjects with different cancers

2017 ◽  
Vol 29 (4) ◽  
pp. 637 ◽  
Author(s):  
Lara Tamburrino ◽  
Marta Cambi ◽  
Sara Marchiani ◽  
Ilaria Manigrasso ◽  
Selene Degl'Innocenti ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cancer Patients ◽  
Dna Fragmentation ◽  
Cell Populations ◽  
Sperm Dna Fragmentation ◽  
Ivf Outcome ◽  
Sperm Cryopreservation ◽  
Reproductive Process ◽  
Sperm Dna

Sperm cryopreservation is widely used by cancer patients undergoing chemo- or radiotherapy. Evidence suggests that IVF outcome with cryopreserved spermatozoa from cancer patients is less successful. To determine whether sperm DNA fragmentation (SDF) is involved in the lower fertilising ability of cryopreserved spermatozoa of cancer patients, SDF was evaluated in thawed spermatozoa from 78 men affected by different cancers and 53 men with non-cancer pathologies. SDF was assessed by the terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL), propidium iodide (PI), flow cytometry procedure, which allows determination of two different cell populations (PIbrighter and PIdimmer) and thus to determine the percentage of DNA fragmented sperm in both. PIdimmer spermatozoa are totally unviable, whereas PIbrighter spermatozoa with SDF may be motile and morphologically normal, having higher biological relevance in the reproductive process. We found that the proportion of DNA fragmented PIbrighter cells was significantly higher in thawed spermatozoa from cancer than non-cancer patients. Moreover, a positive correlation was found between the degree of DNA fragmentation and sperm motility in the PIbrighter population of spermatozoa from cancer patients that wasn’t seen in non-cancer patients. The results of the present study suggest that higher SDF levels may contribute to the lower IVF success of cryopreserved spermatozoa from cancer patients and that evaluation of SDF could complement genetic counselling as part of the routine management of cancer patients who seek fertility preservation.

DETERMINATION OF THE HUMAN SPERM DNA FRAGMENTATION INDEX

Vestnik ◽  
2021 ◽  
pp. 226-232
Author(s):  
К. К. Тлеуханов ◽  
Н. А. Алтыбаева ◽  
М. К. Отарбаев ◽  
Е. М. Тойшибеков ◽  
А. А. Тлеуханова
Keyword(s):  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

В статье представлены собранные данные о методах устранения повышенной частоты фрагментации ДНК у сперматозоидов, которые в некоторых исследованиях подтверждают, что введение антиоксидантов может снизить уровень фрагментации ДНК у сперматозоидов. The article presents collected data on methods of eliminating the increased frequency of DNA fragmentation in spermatozoa, which in some studies confirm that the introduction of antioxidants can reduce the level of DNA fragmentation in spermatozoa.

scholarly journals Critical Aspects of Detection of Sperm DNA Fragmentation by Tunel/Flow Cytometry

2010 ◽  
Vol 56 (4) ◽  
pp. 277-285 ◽  
Author(s):  
Monica Muratori ◽  
Lara Tamburrino ◽  
Sara Marchiani ◽  
Carmen Guido ◽  
Gianni Forti ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Fragmentation ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Critical Aspects

scholarly journals 265 SPERM DNA FRAGMENTATION AND PREGNANCY OUTCOME

2005 ◽  
Vol 17 (2) ◽  
pp. 282
Author(s):  
D. Evenson
Keyword(s):  
Flow Cytometry ◽  
Pregnancy Outcome ◽  
Dna Fragmentation ◽  
Dna Strand Breaks ◽  
Sperm Dna Fragmentation ◽  
Strand Breaks ◽  
Threshold Levels ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Dna Strand

Sperm DNA integrity is obviously important for normal embryo development and pregnancy outcome. Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. The Sperm Chromatin Structure Assay (SCSA) treats sperm with low pH to denature DNA at the sites of DNA strand breaks, followed by acridine orange (AO) staining of green for native DNA and red for denatured DNA, as measured by flow cytometry (FCM), as well as % sperm with high DNA stainability (HDS: immature sperm with intact DNA related to decreased fertilization rates). FCM-sorted sperm from each SCSA-defined population (normal, moderate, and high DNA fragmentation and HDS sperm) show that the moderate DNA fragmentation index (DFI) population has the same image analysis characteristics as normal sperm without significant comets. Thus, an ICSI technician is not likely to differentiate between a normal and a moderate DFI sperm. The TUNEL assay uses an enzyme to add a fluorochrome-labeled base to a 3′-OH broken DNA strand. Both light microscopy and flow cytometry are used for measuring the % and extent of DNA fragmentation but cannot measure the level of HDS. For the COMET assay, sperm are suspended in an electrophoretic gel, placed on a glass microscope slide, digested with proteases and RNAse, subjected to an electric field, and then stained with a DNA dye. The % of comet positive sperm is scored, but the extent of fragmentation is difficult to define and the % HDS cannot be determined. Small pieces of fragmented DNA migrate in the gel forming a “comet.” All three methods have been used for both research and clinical diagnosis and as prognosis for livestock (bulls, boars, rams, stallions) and humans. Light microscope techniques suffer from a lack of statistical soundness needed for clinical decisions as well as present a potential bias in selection of sperm for measurements. Due to the thousands of sperm randomly selected for flow cytometry measurements, the data are statistically robust. Data from all three kinds of measurements in over a hundred manuscripts clearly show that sperm DNA fragmentation has a negative impact on embryo growth and pregnancy. Infertile animals may have nearly all of the sperm with fragmented DNA. Fertility ratings in bulls and boars are clearly related to the percent and extent of DNA fragmentation. Threshold levels for fertile/sub fertile/infertile differ for different species. Likewise different methods/laboratories have suggested various threshold levels to characterize a man with a highly fertile to low/very poor potential. The range of sperm with fragmented DNA is from ∼2% to 100%. The SCSA method has defined a 27–30% DFI as the point in which a man is placed into a statistical category of taking a longer time to achieve in vivo pregnancy, more intrauterine insemination and routine IVF cycles, or no pregnancy. Current data suggest that ICSI may help overcome the diminished pregnancy prognosis with high DFI over the other ART or natural methods.

scholarly journals Obesity as a factor in spermatogenesis disorders (experimental study)

2020 ◽  
Vol 21 (2) ◽  
pp. 36-43
Author(s):  
A. A. Artamonov ◽  
S. V. Bogolyubov ◽  
T. I. Eliseeva ◽  
O. B. Pozdnyakov ◽  
A. V. Astakhova
Keyword(s):  
Total Cholesterol ◽  
Dna Fragmentation ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Study Objective ◽  
Diet Induced Obesity ◽  
White Rats ◽  
Sperm Dna ◽  
Significant Difference

Introduction. In recent years, the effects of obesity on male fertility have been extensively investigated. The results of existing studies are extremely contradictory.The study objective was to determine the effect of obesity on the male reproductive system using the biological model of laboratory rats as an example.Materials and methods. In vivo modeling of diet-induced obesity. The study was conducted on 22 laboratory sexually mature white rats weighing 140–160 g. The animals were divided into two groups: 1 control (10 animals) and 2 rats with diet-induced obesity (12 animals). After 12 weeks, the animals were removed from the experiment. All rats underwent: calculation of the Lee index (body mass index in rats), determination of the concentration and viability of spermatozoa in a suspension of sperm from the epididymis, determination of glucose level of total cholesterol and triglycerides in the blood, study of sperm DNA fragmentation, histological examination testis: calculating the crosssectional area of the seminiferous tubule; determination of the number of non-functioning tubules and tubules with desquamated spermiogenic epithelium; determination of the average spermatogenesis index.Results. In the study groups there were no differences in glucose and total cholesterol levels. However, a statistically significant, significant difference in the level of triglycerides in the blood was revealed. The concentration of sperm and their viability in the studied groups did not differ. The level of sperm DNA fragmentation in the experimental group is significantly higher than in the control group (31.5 ± 10.1 and ± 1.4 %, respectively, p <0.05). Morphometric evaluation of histological preparations did not establish differences in the cross-sectional area of the seminiferous tubules and the average spermatogenesis index in the studied groups. In rats with obesity, compared with the control group, significantly more non-functioning tubules (2.9 ± 0.3 and 8.4 ± 0.3; p <0.05) and tubules with desquamated spermatogenic epithelium (1.8 ± 0.3 and 8.8 ± 0.5; p <0.05).Conclusion. Diet-induced obesity causes impaired spermatogenesis, and damage to the sperm genetic material in male white rats.

scholarly journals Small Variations in Crucial Steps of TUNEL Assay Coupled to Flow Cytometry Greatly Affect Measures of Sperm DNA Fragmentation

2009 ◽  
Vol 31 (4) ◽  
pp. 336-345 ◽  
Author(s):  
M. Muratori ◽  
L. Tamburrino ◽  
V. Tocci ◽  
A. Costantino ◽  
S. Marchiani ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Fragmentation ◽  
Tunel Assay ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

scholarly journals Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Essam-Elden M. Mohammed ◽  
Eman Mosad ◽  
Asmaa M. Zahran ◽  
Diaa A. Hameed ◽  
Emad A. Taha ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Acridine Orange ◽  
Pregnancy Outcome ◽  
Dna Fragmentation ◽  
Chromatin Condensation ◽  
Semen Parameters ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P<0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P<0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

scholarly journals A double-blinded comparison of in situ TUNEL and aniline blue versus flow cytometry acridine orange for the determination of sperm DNA fragmentation and nucleus decondensation state index

Zygote ◽  
2014 ◽  
Vol 23 (4) ◽  
pp. 556-562 ◽  
Author(s):  
Jamal Hamidi ◽  
Christophe Frainais ◽  
Edouard Amar ◽  
Eric Bailly ◽  
Patrice Clément ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Acridine Orange ◽  
Dna Fragmentation ◽  
Aniline Blue ◽  
Sperm Dna Fragmentation ◽  
Acridine Orange Staining ◽  
Sperm Dna ◽  
Double Blinded ◽  
The Impact

SummaryThe impact of sperm DNA fragmentation on assisted reproductive technology (ART) successes, in terms of outcome, is now established. High levels of DNA strand breaks severely affect the probability of pregnancy. The importance of sperm nucleus condensation in early embryogenesis and, subsequently, on the quality of the conceptus is now emerging. In this article we have compared in situ analyses with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) (for DNA fragmentation) with aniline blue (AB) (for nucleus decondensation), versus flow cytometry (FC) after acridine orange staining, in a double-blinded analysis. In our hands, TUNEL and acridine orange give perfectly comparable results. For decondensation the results are also comparable, but the double-stranded green fluorescence obtained with acridine orange seems to slightly underestimate the decondensation status obtained with AB.

Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test

2005 ◽  
Vol 84 (4) ◽  
pp. 833-842 ◽  
Author(s):  
J FERNANDEZ ◽  
L MURIEL ◽  
V GOYANES ◽  
E SEGRELLES ◽  
J GOSALVEZ ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Dispersion Test ◽  
Sperm Chromatin Dispersion Test
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